title: Reconstitution of biochemically altered nuclear pores: transport can be eliminated and restored.
abstract: Biochemically altered nuclear pores specifically lacking the N-acetylglucosamine-bearing pore proteins were constructed in a nuclear assembly extract in order to assign function to these proteins. The depleted pores do not bind nuclear signal sequences or actively import nuclear proteins, but they are functional for diffusion. These defects can be fully repaired by assembly with readded Xenopus pore glycoproteins. Strikingly, isolated rat pore glycoproteins also restore transport. Electron microscopy reveals that depleted pores have largely normal morphology. Thus, the pore glycoproteins are not required for assembly of the nuclear envelope, the major structures of the pore, or a pore diffusional channel. Instead, they are essential for active protein import and, unexpectedly, for construction of the part of the pore necessary for signal sequence recognition.
authors.affiliation: Department of Biology B-022, University of California, San Diego, La Jolla 92093.
authors.forename: D R
authors.initials: DR
authors.lastname: Finlay
authors.forename: D J
authors.initials: DJ
authors.lastname: Forbes
mesh_terms: D000117:Acetylglucosamine / Q000032:analysis; D000818:Animals; D001692:Biological Transport; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005434:Flow Cytometry; D008854:Microscopy, Electron; D009685:Nuclear Envelope / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D011975:Receptors, Mitogen / Q000378:metabolism; D014909:Wheat Germ Agglutinins / Q000378:metabolism; D014981:Xenopus
chemical_list: D009687:Nuclear Proteins; D011975:Receptors, Mitogen; D014909:Wheat Germ Agglutinins; C027503:wheat germ agglutinin receptor; D000117:Acetylglucosamine